Assessment of Radio-Induced Damage in Endothelial Cells Irradiated with 40 kVp, 220 kVp, and 4 MV X-rays by Means of Micro and Nanodosimetric Calculations.
The objective of this work was to study the differences in terms of early biological effects that might exist between different X-rays energies by using a mechanistic approach. To this end, radiobiological experiments exposing cell monolayers to three X-ray energies were performed in order to assess the yields of early DNA damage, in particular of double-strand breaks (DSBs). The simulation of these irradiations was set in order to understand the differences in the obtained experimental results. Hence, simulated results in terms of microdosimetric spectra and early DSB induction were analyzed and compared to the experimental data. Human umbilical vein endothelial cells (HUVECs) were irradiated with 40, 220 kVp, and 4 MV X-rays. The Geant4 Monte Carlo simulation toolkit and its extension Geant4-DNA were used for the simulations. Microdosimetric calculations aiming to determine possible differences in the variability of the energy absorbed by the irradiated cell population for those photon spectra were performed on 10,000 endothelial cell nuclei representing a cell monolayer. Nanodosimetric simulations were also carried out using a computation chain that allowed the simulation of physical, physico-chemical, and chemical stages on a single realistic endothelial cell nucleus model including both heterochromatin and euchromatin. DNA damage was scored in terms of yields of prompt DSBs per Gray (Gy) and per giga (109) base pair (Gbp) and DSB complexity was derived in order to be compared to experimental data expressed as numbers of histone variant H2AX (γ-H2AX) foci per cell. The calculated microdosimetric spread in the irradiated cell population was similar when comparing between 40 and 220 kVp X-rays and higher when comparing with 4 MV X-rays. Simulated yields of induced DSB/Gy/Gbp were found to be equivalent to those for 40 and 220 kVp but larger than those for 4 MV, resulting in a relative biological effectiveness (RBE) of 1.3. Additionally, DSB complexity was similar between the considered photon spectra. Simulated results were in good agreement with experimental data obtained by IRSN (Institut de radioprotection et de sûreté nucléaire) radiobiologists. Despite differences in photon energy, few differences were observed when comparing between 40 and 220 kVp X-rays in microdosimetric and nanodosimetric calculations. Nevertheless, variations were observed when comparing between 40/220 kVp and 4 MV X-rays. Thanks to the simulation results, these variations were able to be explained by the differences in the production of secondary electrons with energies below 10 keV.


Novel laser-processed CsI:Tl detector for SPECT.
The aim of this work is to demonstrate the feasibility of a novel technique for fabrication of high spatial resolution CsI:Tl scintillation detectors for single photon emission computed tomography systems. The scintillators are fabricated using laser-induced optical barriers technique to create optical microstructures (or optical barriers) inside the CsI:Tl crystal bulk. The laser-processed CsI:Tl crystals are 3, 5, and 10 mm in thickness. In this work, the authors focus on the simplest pattern of optical barriers in that the barriers are created in the crystal bulk to form pixel-like patterns resembling mechanically pixelated scintillators. The monolithic CsI:Tl scintillator samples are fabricated with optical barrier patterns with 1.0 × 1.0 mm(2) and 0.625 × 0.625 mm(2) pixels. Experiments were conducted to characterize the fabricated arrays in terms of pixel separation and energy resolution. A 4 × 4 array of multipixel photon counter was used to collect the scintillation light in all the experiments. The process yield for fabricating the CsI:Tl arrays is 100% with processing time under 50 min. From the flood maps of the fabricated detectors exposed to 122 keV gammas, peak-to-valley (P/V) ratios of greater than 2.3 are calculated. The P/V values suggest that regardless of the crystal thickness, the pixels can be resolved. The results suggest that optical barriers can be considered as a robust alternative to mechanically pixelated arrays and can provide high spatial resolution while maintaining the sensitivity in a high-throughput and cost-effective manner.


[Demodex-related marginal blepharitis in Japan].
Some marginal blepharitis is related to demodex, but this has not yet been reported in Japan. In this study, patients with severe marginal blepharitis with cylindrical dandruff were studied to examine the number of demodex in their cilia. Ten eyes of 10 patients (7 men and 3 women, mean age: 62.9 +/- 9.0 years) with unilateral marginal blepharitis which had cylindrical dandruff in their cilia were studied. Three cilia which had the most cylindrical dandruff were removed from the eyelids for microscopic examination. After cleaning the eyelid margins for 1 week, the examination was repeated. Scoring of itching and foreign body sensation of the patients was performed both before and after the treatment. Demodex folliculorum was detected in the cilia of 8 out of 10 eyes (80%), and 22 cilia out of 30 with cylindrical dandruff. The average number of demodex/cilia was 1.6 +/- 0.9. In all cases, the number of demodex/cillium decreased significantly with the improvement in symptoms and blepharitis. An Increase in the number of demodex might be the pathogen causing blepharitis with cylindrical dandruff, and cleaning of the eyelid margin is effective as a therapeutic method.


Quantifying the Subharmonic Mucosal Wave in Excised Larynges via Digital Kymography.
In this paper, a nonlinear least squares fitting method was proposed to quantify subharmonic mucosal waves. Subharmonic mucosal waves from 10 excised canine larynges were recorded using digital kymography and analyzed using nonlinear least squares and linear least squares methods. Amplitudes of fundamental and subharmonic mucosal wave components of right-upper, right-lower, left-upper, and left-lower vocal fold lips were calculated. Lastly, phase differences of fundamental and subharmonic components of the left and right vocal folds were compared. The results showed that the nonlinear least squares analysis method provides a more effective complement to the linear fitting method for subharmonic mucosal wave extraction. There was a significant difference in amplitudes between the subharmonic and the fundamental components of mucosal waves (P < 0.05). The phase differences of the fundamental and the subharmonic components of the right and left vocal folds were not significantly different. The application of the nonlinear least squares analysis method in digital kymography is useful for the characterization of subharmonic mucosal waves.


[Effectiveness of methotrexate for the escape by salazosulfapyridine].
Although disease modifying anti-rheumatic drugs (DMARDs) are used in the treatment of rheumatoid arthritis (RA), the selection of agents in the case of relapse (escape phenomenon) lacks clear-cut standards. We compared the effectiveness in a salazosulfapyridine and then methotrexate (SASP-->MTX) group with that in the mothotrexate (SASP+MTX) group after escape phenomenon expression in C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) data. Outpatients of the Matsubara Mayflower Hospital with a history of DMARD administration during the 4 years prior to May 2003 were studied. The CRP level in the SASP-->MTX group (n=8) after the escape phenomenon expression showed a decline after 3 months, but no decline was seen even after 3 months the two in the CRP level in the SASP+MTX group (n=10). However, the difference between groups was not significant. The fluctuation in ESR was similar to that in CRP. However, ESR was significantly lower in the SASP-->MTX group 20 weeks after escape phenomenon expression. In evaluating treatment effectiveness after escape phenomenon expression in each group, SASP-->MTX was effective in 10 and SASP+MTX in 7 patients. Side effects necessitated cessation of treatment in 1 patient in the SASP-->MTX group. Treatment continued in 4 patients in the SASP-->MTX group and 2 in the SASP+MTX group, even though side effects occurred. It should be borne in mind that combination therapy often has greater clinical benefit than single agent therapy but not always.


[Nature of autobiographical memory that facilitates improvement of negative moods].
Persons in negative moods often recall their positive experiences to alleviate their affective states. This study investigated the nature of autobiographical memories that facilitate improvement of negative moods. More specifically, it examined the relations between participants' subjective moods and the three natures of recalled memories (the current positivity, the past positivity, and the importance). After 67 participants were induced either negative or neutral moods, they were asked to recall five positive experiences and to rate their affective states. Results indicated that the more important memories participants recalled, the more positive moods they reported after recollection, and that these effects of the importance of autobiographical memories were observed in both negative and neutral moods. In contrast, neither the current positivity of memories nor the past positivity of memories significantly influenced participant's moods in both negative and neutral moods. The mechanism by which the importance of autobiographical memory affects moods was discussed.


The changing face of HIV vaccine research.
While there has been remarkable progress in understanding the biology of HIV-1 and its recognition by the human immune system, we have not yet developed an efficacious HIV-1 vaccine. Vaccine challenges include the genetic diversity and mutability of HIV-1 which create a plethora of constantly changing antigens, the structural features of the viral envelope glycoprotein that disguise conserved receptor-binding sites from the immune system, and the presence of carbohydrate moieties that shield potential epitopes from antibodies. Despite these challenges, there has been significant scientific progress in recent years. In 2009, a large-scale clinical trial known as RV144 demonstrated that a HIV-1 vaccine could modestly reduce the incidence of HIV-1 infection. Further, the identification of broadly neutralizing monoclonal antibodies (such as VRC01, a human monoclonal antibody capable of neutralizing over 90% of natural HIV-1 isolates, as well as PG and PGT antibodies that recognize conserved glycopeptide epitopes) has revealed new opportunities for vaccine design. Our ability to understand HIV-1 structure and antibody epitopes at the atomic level, the rapid advance of computational and bioinformatics approaches to immunogen design, and our newly acquired knowledge that it is possible for a vaccine to reduce the risk of HIV-1 infection, have all opened up new and promising pathways towards the development of an urgently needed effective HIV-1 vaccine. This article summarizes challenges to the development of an HIV-1 vaccine, lessons learned from scientific investigation and completed vaccine trials, and promising developments in HIV-1 vaccine design.


[Effects of electroacupuncture on resting-state encephalic functional connectivity network in patients with PTSD].
To explore the central regulatory mechanism of electroacupuncture (EA) on patients with post-traumatic stress disorder (PTSD). Fourteen patients of PTSD were selected as study objects and treated with "regulating mind and restoring consciousness" acupuncture method, in which Baihui (GV 20) and Shenting (GV 24) were used as main acupoints and Sishencong (EX-HN 1) and Fengchi (GB 20) were used as supporting acupoints for acupuncture. After the arrival of qi, Han's acupoint nerve stimulator was connected for 30 min per treatment, three times a week for consecutive 12 weeks. Before treatment and 12 weeks into treatment, the clinician administered PTSD scale (CAPS), self-rating anxiety scale (SAS) and self-rating depression scale (SDS) were evaluated; a Siemens 3.0 T magnetic resonance imaging system was used to perform resting-state scan, and bilateral hippocampus were taken as region of interested to perform encephalic function connectivity analysis. After the treatment, the scores of CAPS, SAS and SDS were all reduced compared with those before treatment (all P<0.05) ; function connectivity was enhanced in bilateral hippocampus, right posterior central gyrus and left superior parietal lobule (2.3<Z<4.0, P<0.05), but the connectivity in left parahippocampal gyrus and right amygdaloid nucleus was suppressed. Electroacupuncture has certain improving effects on PTSD symptoms, which is likely to be related with enhancing the connectivity between parietal lobe and hippocampus, suppressing the connectivity between hippocampus and parahippocampal gyrus, amygdaloid, leading to an indirect influence on limbic system.


Natural infections of Anopheles albimanus with Plasmodium in a small malaria focus.
Entomologic surveys conducted in a small village in an area of known high malaria transmission in El Salvador yielded a high rate of infection in Anopheles albimanus collected inside houses in which cases of malaria had occurred. Of 324 specimens dissected, 12 were found to harbor sporozoites or oocysts. This is in contrast to prior reports of extremely low infection rates in collections of this species from malarious areas, and suggests that under some circumstances A. albimanus does meet the criteria of an effective malaria vector.


Puddles, parties, and professors: linking word categorization to neural patterns of visuospatial coding.
Behavioral evidence suggests that during word processing people spontaneously map object, valence, and power information to locations in vertical space. Specifically, whereas "overhead" (e.g., attic), positive (e.g., party), and powerful nouns (e.g., professor) are associated with "up," "underfoot" (e.g., carpet), negative (e.g., accident), and powerless nouns (e.g., assistant) are associated with "down." What has yet to be elucidated, however, is the precise nature of these effects. To explore this issue, an fMRI experiment was undertaken, during which participants were required to categorize the position in which geometrical shapes appeared on a computer screen (i.e., upper or lower part of the display). In addition, they also judged a series of words with regard to location (i.e., up vs. down), valence (i.e., good vs. bad), and power (i.e., powerful vs. powerless). Using multivoxel pattern analysis, it was found that classifiers that successfully distinguished between the positions of shapes in subregions of the inferior parietal lobe also provided discriminatory information to separate location and valence, but not power word judgments. Correlational analyses further revealed that, for location words, pattern transfer was more successful the stronger was participants' propensity to use visual imagery. These findings indicate that visual coding and conceptual processing can elicit common representations of verticality but that divergent mechanisms may support the reported effects.


Essential role of the C-terminus in Melanocarpus albomyces laccase for enzyme production, catalytic properties and structure.
The C-terminus of the fungal laccase from Melanocarpus albomyces (MaL) is processed during secretion at a processing site conserved among the ascomycete laccases. The three-dimensional structure of MaL has been solved as one of the first complete laccase structures. According to the crystal structure of MaL, the four C-terminal amino acids of the mature protein penetrate into a tunnel leading towards the trinuclear site. The C-terminal carboxylate group forms a hydrogen bond with a side chain of His140, which also coordinates to the type 3 copper. In order to analyze the role of the processed C-terminus, site-directed mutagenesis of the MaL cDNA was performed, and the mutated proteins were expressed in Trichoderma reesei and Saccharomyces cerevisiae. Changes in the C-terminus of MaL caused major defects in protein production in both expression hosts. The deletion of the last four amino acids dramatically affected the activity of the enzyme, as the deletion mutant delDSGL(559) was practically inactive. Detailed characterization of the purified L559A mutant expressed in S. cerevisiae showed the importance of the C-terminal plug for laccase activity, stability, and kinetics. Moreover, the crystal structure of the L559A mutant expressed in S. cerevisiae showed that the C-terminal mutation had clearly affected the trinuclear site geometry. The results in this study clearly confirm the critical role of the last amino acids in the C-terminus of MaL.


Lexical Preactivation in Basic Linguistic Phrases.
Many previous studies have shown that predictable words are read faster and lead to reduced neural activation, consistent with a model of reading in which words are activated in advance of being encountered. The nature of such preactivation, however, has typically been studied indirectly through its subsequent effect on word recognition. Here, we use magnetoencephalography to study the dynamics of prediction within serially presented adjective-noun phrases, beginning at the point at which the predictive information is first available to the reader. Using corpus transitional probability to estimate the predictability of a noun, we found an increase in activity in the left middle temporal gyrus in response to the presentation of highly predictive adjectives (i.e., adjectives that license a strong noun prediction). Moreover, we found that adjective predictivity and expected noun frequency interacted, such that the response to the highly predictive adjectives (e.g., stainless) was modulated by the frequency of the expected noun (steel). These results likely reflect preactivation of nouns in highly predictive contexts. The fact that the preactivation process was modulated by the frequency of the predicted item is argued to provide support for a frequency-sensitive lexicon.


The role of infectious disease, inbreeding and mating preferences in maintaining MHC genetic diversity: an experimental test.
In house mice, and probably most mammals, major histocompatibility complex (MHC) gene products influence both immune recognition and individual odours in an allele-specific fashion. Although it is generally assumed that some form of pathogen-driven balancing selection is responsible for the unprecedented genetic diversity of MHC genes, the MHC-based mating preferences observed in house mice are sufficient to account for the genetic diversity of MHC genes found in this and other vertebrates. These MHC disassortative mating preferences are completely consistent with the conventional view that pathogen-driven MHC heterozygote advantage operates on MHC genes. This is because such matings preferentially produce MHC-heterozygours progeny, which could enjoy enhanced disease resistance. However, such matings could also function to avoid genome-wide inbreeding. To discriminate between these two hypotheses we measured the fitness consequences of both experimentally manipulated levels of inbreeding and MHC homozygosity and heterozygosity in semi-natural populations of wild-derived house mice. We were able to measure a fitness decline associated with inbreeding, but were unable to detect fitness declines associated with MHC homozygosity. These data suggest that inbreeding avoidance may be the most important function of MHC-based mating preferences and therefore the fundamental selective force diversifying MHC genes in species with such mating patterns. Although controversial, this conclusion is consistent with the majority of the data from the inbreeding and immunological literature.


Doctors' perspectives on adhering to advance care directives when making medical decisions for patients: an Australian interview study.
Advance care planning (ACP) assists people to identify their goals, values and treatment preferences for future care. Ideally, preferences are documented in an advance care directive (ACD) and used by doctors to guide medical decision-making should the patient subsequently lose their decision-making capacity. However, studies demonstrate that ACDs are not always adhered to by doctors in clinical practice. We aim to describe the attitudes and perspectives of doctors regarding ACD adherence and the utility of ACDs in clinical practice. Face-to-face semistructured interviews were conducted using three case-based vignettes to explore doctors' decision-making and attitudes towards ACDs. Transcripts were analysed using a thematic analysis. Doctors from a variety of medical specialties and with varying experience levels were recruited from a large tertiary hospital in Melbourne, Australia. A total of 21 doctors were interviewed, 48% female (10/21). Most (19/21) reported having experience using ACDs. Four themes were identified: aligning with patient preferences (avoiding unwanted care, prioritising autonomy and anticipating family opposition), advocating best interests (defining futile care, relying on clinical judgement, rejecting unreasonable decisions and disregarding legal consequences), establishing validity (doubting rigour of the decision-making process, questioning patients' ability to understand treatment decisions, distrusting outdated preferences and seeking confirmation) and translating written preferences into practice (contextualising patient preferences, applying subjective terminology and prioritising emergency medical treatment). ACDs provide doctors with opportunities to align patient preferences with treatment and uphold patient autonomy. However, doctors experience decisional conflict when attempting to adhere to ACDs in practice, especially when they believe that adhering to the ACD is not in the patients' best interests, or if they doubt the validity of the ACD. Future ACP programmes should consider approaches to improve the validity and applicability of ACDs. In addition, there is a need for ethical and legal education to support doctors' knowledge and confidence in ACP and enacting ACDs.


Changes in light intensity reveal a major role for carbon balance in Arabidopsis responses to high temperature.
High temperature (HT) is a major limiting factor for plant productivity. Because some responses to HT, notably hyponasty, resemble those encountered in low light (LL), we hypothesized that plant responses to HT are under the control of carbon balance. We analysed the interactive effects of HT and irradiance level on hyponasty and a set of traits related to plant growth in natural accessions of Arabidopsis thaliana and mutants affected in heat dissipation through transpiration (NCED6-OE, ost2) and starch metabolism (pgm). HT induced hyponasty, reduced plant growth and modified leaf structure. LL worsened the effects of HT, while increasing light restored trait values close to levels observed at control temperature. Leaf temperature per se did not play a major role in the observed responses. By contrast, a major role of carbon balance was supported by hyponastic growth of pgm, as well as morphological, physiological (photosynthesis, sugar and starch contents) and transcriptional data. Carbon balance could be a common sensor of HT and LL, leading to responses specific of the shade avoidance syndrome. Hyponasty and associated changes in plant traits could be key traits conditioning plant performance under competition for light, particularly in warm environments.


3D visual activity assessment based on natural scene statistics.
One of the most challenging ongoing issues in the field of 3D visual research is how to perceptually quantify object and surface visualizations that are displayed within a virtual 3D space between a human eye and 3D display. To seek an effective method of quantification, it is necessary to measure various elements related to the perception of 3D objects at different depths. We propose a new framework for quantifying 3D visual information that we call 3D visual activity (3DVA), which utilizes natural scene statistics measured over 3D visual coordinates. We account for important aspects of 3D perception by carrying out a 3D coordinate transform reflecting the nonuniform sampling resolution of the eye and the process of stereoscopic fusion. The 3DVA utilizes the empirical distortions of wavelet coefficients to a parametric generalized Gaussian probability distribution model and a set of 3D perceptual weights. We conducted a series of simulations that demonstrate the effectiveness of the 3DVA for quantifying the statistical dynamics of visual 3D space with respect to disparity, motion, texture, and color. A successful example application is also provided, whereby 3DVA is applied to the problem of predicting visual fatigue experienced when viewing 3D displays.


Prevention of mother-to-child transmission of HIV Option B+ cascade in rural Tanzania: The One Stop Clinic model.
Strategies to improve the uptake of Prevention of Mother-To-Child Transmission of HIV (PMTCT) are needed. We integrated HIV and maternal, newborn and child health services in a One Stop Clinic to improve the PMTCT cascade in a rural Tanzanian setting. The One Stop Clinic of Ifakara offers integral care to HIV-infected pregnant women and their families at one single place and time. All pregnant women and HIV-exposed infants attended during the first year of Option B+ implementation (04/2014-03/2015) were included. PMTCT was assessed at the antenatal clinic (ANC), HIV care and labour ward, and compared with the pre-B+ period. We also characterised HIV-infected pregnant women and evaluated the MTCT rate. 1,579 women attended the ANC. Seven (0.4%) were known to be HIV-infected. Of the remainder, 98.5% (1,548/1,572) were offered an HIV test, 94% (1,456/1,548) accepted and 38 (2.6%) tested HIV-positive. 51 were re-screened for HIV during late pregnancy and one had seroconverted. The HIV prevalence at the ANC was 3.1% (46/1,463). Of the 39 newly diagnosed women, 35 (90%) were linked to care. HIV test was offered to >98% of ANC clients during both the pre- and post-B+ periods. During the post-B+ period, test acceptance (94% versus 90.5%, p<0.0001) and linkage to care (90% versus 26%, p<0.0001) increased. Ten additional women diagnosed outside the ANC were linked to care. 82% (37/45) of these newly-enrolled women started antiretroviral treatment (ART). After a median time of 17 months, 27% (12/45) were lost to follow-up. 79 women under HIV care became pregnant and all received ART. After a median follow-up time of 19 months, 6% (5/79) had been lost. 5,727 women delivered at the hospital, 20% (1,155/5,727) had unknown HIV serostatus. Of these, 30% (345/1,155) were tested for HIV, and 18/345 (5.2%) were HIV-positive. Compared to the pre-B+ period more women were tested during labour (30% versus 2.4%, p<0.0001). During the study, the MTCT rate was 2.2%. The implementation of Option B+ through an integrated service delivery model resulted in universal HIV testing in the ANC, high rates of linkage to care, and MTCT below the elimination threshold. However, HIV testing in late pregnancy and labour, and retention during early ART need to be improved.


Differential contribution of genomic regions to marked genetic variation and prediction of quantitative traits in broiler chickens.
Genome-wide association studies in humans have found enrichment of trait-associated single nucleotide polymorphisms (SNPs) in coding regions of the genome and depletion of these in intergenic regions. However, a recent release of the ENCyclopedia of DNA elements showed that ~80 % of the human genome has a biochemical function. Similar studies on the chicken genome are lacking, thus assessing the relative contribution of its genic and non-genic regions to variation is relevant for biological studies and genetic improvement of chicken populations. A dataset including 1351 birds that were genotyped with the 600K Affymetrix platform was used. We partitioned SNPs according to genome annotation data into six classes to characterize the relative contribution of genic and non-genic regions to genetic variation as well as their predictive power using all available quality-filtered SNPs. Target traits were body weight, ultrasound measurement of breast muscle and hen house egg production in broiler chickens. Six genomic regions were considered: intergenic regions, introns, missense, synonymous, 5' and 3' untranslated regions, and regions that are located 5 kb upstream and downstream of coding genes. Genomic relationship matrices were constructed for each genomic region and fitted in the models, separately or simultaneously. Kernel-based ridge regression was used to estimate variance components and assess predictive ability. Contribution of each class of genomic regions to dominance variance was also considered. Variance component estimates indicated that all genomic regions contributed to marked additive genetic variation and that the class of synonymous regions tended to have the greatest contribution. The marked dominance genetic variation explained by each class of genomic regions was similar and negligible (~0.05). In terms of prediction mean-square error, the whole-genome approach showed the best predictive ability. All genic and non-genic regions contributed to phenotypic variation for the three traits studied. Overall, the contribution of additive genetic variance to the total genetic variance was much greater than that of dominance variance. Our results show that all genomic regions are important for the prediction of the targeted traits, and the whole-genome approach was reaffirmed as the best tool for genome-enabled prediction of quantitative traits.


Self-care and Its Predictors in Patients With Chronic Heart Failure in Western Iran.
Chronic heart failure (CHF) is an increasing and costly health problem worldwide. Effective self-care behaviors reduce the cost and improve CHF outcomes. Interventions targeting improvement of self-care need to identify the baseline status of patients and factors associated with self-care to tailor the programs to patients' needs. The aim of this study was to describe self-care and its predictors in patients with CHF in western Iran. In a cross-sectional study, 255 patients with CHF in Kermanshah were recruited and 231 (mean [SD] age, 66 [13] years; 51.5% women) completed the interviews. Self-care maintenance, self-care management, and self-care confidence were evaluated using a Persian heart failure self-care index. Each of these 3 measures had a total possible score of 100, with 22 indicators. The mean (SD) self-care scores were low: maintenance, 33.8 (10.7); management, 32.2 (12.0); and confidence, 43.6 (15.6). Self-care maintenance was significantly and positively associated with education, disease duration, and living conditions. Self-care management was significantly and positively associated with education and number of hospital admissions. However, the parameter estimates in all those relationships were small. Self-care in patients with CHF in Iran needs major improvement, and many determinants of self-care identified by other studies were not consistently associated with poor self-care scores in Iran. Further research considering a wide range of factors associated with self-care (eg, socioeconomic and health system-related factors) and application of culturally relevant interventional strategies is recommended.


Metabolic profiling reveals disorder of amino acid metabolism in four brain regions from a rat model of chronic unpredictable mild stress.
Chronic stress is closely linked to clinical depression, which could be assessed by a chronic unpredictable mild stress (CUMS) animal model. We present here a GC/MS-based metabolic profiling approach to investigate neurochemical changes in the cerebral cortex, hippocampus, thalamus, and remaining brain tissues. Multi-criteria assessment for multivariate statistics could identify differential metabolites between the CUMS-model rats versus the healthy controls. This study demonstrates that the significantly perturbed metabolites mainly involving amino acids play an indispensable role in regulating neural activity in the brain. Therefore, results obtained from such metabolic profiling strategy potentially provide a unique perspective on molecular mechanisms of chronic stress.


A selenocysteine variant of the human copper chaperone for superoxide dismutase. A Se-XAS probe of cluster composition at the domain 3-domain 3 dimer interface.
We report the semisynthesis of a selenocysteine (Sec) derivative of the human copper chaperone for superoxide dismutase, substituted with Sec at the C-terminal C246 residue. Measurements of hCCS-induced SOD1 activation were used to show that the C-terminal CXC sequence is both necessary and sufficient for EZn-SOD maturation. Therefore, an active CAU variant carrying Sec as the terminal amino acid was prepared by expressed protein ligation of a single selenocysteine amino acid to a 243-CA truncation. This reaction proceeded in high yield and generated the desired 243-CAX (X = C or U) protein with the expected mass. Se-edge XAS of the apoprotein indicated that both Se-S and Se-Se interactions were present in a 0.3:0.7 ratio, indicating an equilibrium between species with either a selenosulfide or a diselenide cross-link. After reduction on immobilized TCEP, the ligated Cys and Sec apoproteins bound up to 2.5 Cu(I) ions per hCCS monomer with both Cu and Se as constituent atoms of the cluster which forms at the domain 3 interface of a hCCS dimer. Merging of XAS data at the Cu and Se K-absorption edges provided additional details of the cluster composition, specifically the fact that both Se atoms occupied bridging positions between two Cu(I) atoms. Further, the requirement for identical Cu-Se bond lengths and Debye-Waller factors at each absorption edge allowed us to rule out simple models for the cluster composition such as a bis-Cys(Sec)-bridged dinuclear cluster and was indicative of a more complex cluster with a nuclearity of >or=3.


Smac induces cytochrome c release and apoptosis independently from Bax/Bcl-x(L) in a strictly caspase-3-dependent manner in human carcinoma cells.
The mitochondrial apoptosis pathway mediates cell death through the release of various pro-apoptotic factors including cytochrome c and Smac, the second mitochondrial activator of caspases, into the cytosol. Smac was shown previously to inhibit IAP proteins and to facilitate initiation of the caspase cascade upon cytochrome c release. To investigate Smac function during apoptosis and to explore Smac as an experimental cancer therapeutic, we constructed an expression system based on a single adenoviral vector containing Smac under control of the Tet-off system supplied in cis. Conditional expression of Smac induced apoptosis in human HCT116 and DU145 carcinoma cells regardless of the loss of Bax or overexpression of Bcl-x(L). Nevertheless, apoptosis induced by Smac was associated with cytochrome c release and breakdown of the mitochondrial membrane potential. This indicates that Smac acts independently of Bax and Bcl-x(L) during initiation of apoptosis and triggers a positive feedback loop that results in Bax/Bcl-x(L)-independent activation of mitochondria. In caspase-proficient cells, Smac-induced apoptosis could be inhibited partially by cell-permeable LEHD (caspase-9 inhibitor) and DEVD (caspase-3 inhibitor) peptides. Furthermore, loss of caspase-3 expression in MCF-7 cells carrying a caspase-3 null mutation completely abrogated the sensitivity for Smac-induced apoptotic or nonapoptotic, necrosis-like cell death, while re-expression of caspase-3 conferred sensitivity. Altogether, caspase-3 but not caspase-9 activation was necessary for execution of Smac-induced cell death. Notably, Smac did not induce caspase-9 processing in the absence of caspase-3. Thus, caspase-9 processing occurs secondary to caspase-3 activation during Smac-induced apoptosis. Altogether, Smac is capable of circumventing defects in mitochondrial apoptosis signaling such as loss of Bax or overexpression of Bcl-x(L) that are frequently observed in tumor cells resistant to anticancer therapy. Consequently, Smac appears to be a promising therapeutic target in anticancer treatment.


Loss of color vision and Stiles' II1 mechanism in a patient with cerebral infarction.
A 70-year old man developed achromatopsia with bilateral loss of superior visual fields and an inability to recognize familiar faces (prosopagnosia). Ophthalmologic examination results were normal. Visual acuity was 20/25 in either eye. Computerized axial tomography of the brain revealed infarction of the inferior aspect of the temporal occipital cortex in both hemispheres. The patient's complaint that objects appeared only in shades of gray was supported by large errors made throughout the spectrum on the Farnsworth-Munsell 100 hue test and by matches over the entire red/green range on the Nagel anomaloscope. Although absolute scotopic and photopic thresholds were unremarkable, the increment thresholds to a 482-nm test on a red background increased monotonically as if the II1 mechanism were absent. In addition, the spectral sensitivity to large test flashes on an intense red background peaked in the middle rather than in the short-wave portion of the spectrum, as is normally found. We speculate that the chromatic channel is compromised. The patient's residual vision is mediated by a luminance channel that is subserved by the middle and long--but not the short--wave cone mechanisms.


Intravenous lidocaine decreases tumor necrosis factor alpha expression both locally and systemically in pigs undergoing lung resection surgery.
Lung resection surgery is associated with an inflammatory reaction. The use of 1-lung ventilation (OLV) seems to increase the likelihood of this reaction. Different prophylactic and therapeutic measures have been investigated to prevent lung injury secondary to OLV. Lidocaine, a commonly used local anesthetic drug, has antiinflammatory activity. Our main goal in this study was to investigate the effect of IV lidocaine on tumor necrosis factor α (TNF-α) lung expression during lung resection surgery with OLV. Eighteen pigs underwent left caudal lobectomy. The animals were divided into 3 groups: control, lidocaine, and sham. All animals received general anesthesia. In addition, animals in the lidocaine group received a continuous IV infusion of lidocaine during surgery (1.5 mg/kg/h). Animals in the sham group only underwent thoracotomy. Samples of bronchoalveolar lavage (BAL) fluid and plasma were collected before initiation of OLV, at the end of OLV, at the end of surgery, and 24 hours after surgery. Lung biopsy specimens were collected from the left caudal lobe (baseline) before surgery and from the mediastinal lobe and the left cranial lobe 24 hours after surgery. Samples were flash-frozen and stored to measure levels of the following inflammatory markers: interleukin (IL) 1β, IL-2, IL-10, TNF-α, nuclear factor κB, monocyte chemoattractant protein-1, inducible nitric oxide synthase, and endothelial nitric oxide synthase. Markers of apoptosis (caspase 3, caspase 9, Bad, Bax, and Bcl-2) were also measured. In addition, levels of metalloproteinases and nitric oxide metabolites were determined in BAL fluid and in plasma samples. A nonparametric test was used to examine statistical significance. OLV caused lung damage with increased TNF-α expression in BAL, plasma, and lung samples. Other inflammatory (IL-1β, nuclear factor κB, monocyte chemoattractant protein-1) and apoptosis (caspase 3, caspase 9, and BAX) markers were also increased. With the use of IV lidocaine there was a significant decrease in the levels of TNF-α in the same samples compared with the control group. Lidocaine administration also reduced the inflammatory and apoptotic changes observed in the control group. Hemodynamic values, blood gas values, and airway pressure were similar in all groups. Our results suggest that lidocaine can prevent OLV-induced lung injury through reduced expression of proinflammatory cytokines and lung apoptosis. Administration of lidocaine may help to prevent lung injury during lung surgery with OLV.


Adenosine 3',5'-monophosphate content in rat caudate nucleus: demonstration of dopaminergic and adrenergic receptors.
Dopamine, apomorphine, isoproterenol, and norepinephrine each increased the concentration of adenosine 3',5'-monophosphate in slice of rat caudate nucleus. The concentrations of dopamine, apomorphine isoproterenol, and norepinephrine causing half-maximal increases were 60, 150, 0.03 and 30 micromoles per liter, respectively. The effect of dopamine was blocked by fluphenazine, a dopamine receptor antagonist, but not by propranolol, a beta-andrenergic receptor antagonist. Conversely, the effect of isoproterenol was blocked by propranolol but not by fluphenazine. The results suggest that in rat caudate nucleus there are two distinct catecholamine receptors capable of causing increased concentrations of adenosine 3',5'-monophosphate, one having the characteristic of dopamine receptor, and the other having the characteristics of beta-adrenergic receptor.


Caspase inhibitors reduce the apoptotic but not necrotic component of kainate injury in primary murine cortical neuronal cultures.
Excitotoxicity has been demonstrated to play a major role in ischemic neuronal injury. While the necrotic component of excitotoxicity has been well demonstrated, apoptosis has also been shown to play a role. We sought to quantitate and modulate the apoptotic component of kainate-induced injury. Experiments were performed in mouse primary cortical neuronal cultures after three or 10 days in vitro. Cell death was assessed by Hoechst/propidium iodide staining and cell counting. Apoptosis was further confirmed with inhibition by caspase inhibitors. Exposure of three-day old neurons to 100 microM kainate produced an injury in which 56% +/- 0.9% of cells showed apoptotic nuclei and 13.5% +/- 2.0% showed necrotic nuclei. After 10 days in vitro neurons were more easily injured by kainate, but the cell death had primarily necrotic characteristics. Inhibition of both caspases 1 and 3 significantly reduced the apoptotic injury of 3-day old neurons. Neither reduced the necrotic component. Inhibition of protein synthesis with cycloheximide was also effective in reducing the apoptotic injury without affecting the necrotic injury. Kainate injury causes both apoptosis and necrosis, with the injury depending on both the dose of kainate and the age of the culture. The apoptotic component can be selectively reduced by caspase inhibition or cycloheximide.


Benthic marine debris in the Bay of Fundy, eastern Canada: Spatial distribution and categorization using seafloor video footage.
Marine debris, particularly plastic and abandoned, lost and discarded fishing gear, is ubiquitous in marine environments. This study provides the first quantitative and qualitative assessment of benthic debris using seafloor video collected from a drop camera system in the Bay of Fundy, Eastern Canada. An estimated 137 debris items km-2 of seafloor were counted, comprising of plastic (51%), fishing gear (including plastic categories; 28%) and other (cable, metal, tires; 21%). Debris was widespread, but mainly located nearshore (within 9 km) and on the periphery of areas with high fishing intensity. This baseline benthic marine debris characterization and estimate of abundance provides valuable information for government (municipal, provincial and federal) and for other stakeholders to implement management strategies to reduce plastic and other categories of benthic marine pollution at source. Strategies may include limiting plastic use and reducing illegal dumping through improved education among fishers.


[Genetic diagnosis and molecular pathology of inherited neuropathy].
Recent advances in genetic analysis technology have enabled a surprising progress in genetic diagnosis in the field of neurological disease research. High-throughput molecular biology techniques, such as microarrays and next-generation sequencing, are the major contributors to this progress and to new discoveries. Charcot-Marie-Tooth disease (CMT), a known hereditary motor and sensory neuropathy, is clinically and genetically heterogeneous. Genetic studies have revealed at least 35 disease causing-genes responsible for Charcot-Marie-Tooth disease. Genetic studies have revealed that abnormalities in the following factors are the cause of inherited neuropathies: myelin components, transcription factors controlling myelination, myelin maintenance system, differentiation factors related to the peripheral nerve, neurofilaments, protein transfer system, mitochondrial proteins, DNA repair, RNA/protein synthesis, ion channels, and aminoacyl-tRNA synthetase. On the other hand concomitant with the increase in the number of genes that must be screened for mutations, the labor and reagent costs for molecular genetic testing have increased significantly. Therefore, new methodology for detecting gene mutations is required. Based on the recent progress in DNA analysis methods, resequencing microarray appears to be an economical and highly sensitive method for detecting mutations. We have been screening CMT patients for mutations using originally designed microarray DNA chips since 2007, thencehaving identified disease causing mutations in MPZ, GJB1, PMP22, EGR2, MFN2, NEFL, PRX, AARS, GARS, DNM2, and SETX genes in CMT patients.


Pest categorisation of beet necrotic yellow vein virus.
Following a request from the EU Commission, the Panel on Plant Health performed a categorisation of beet necrotic yellow vein virus (BNYVV), the causal agent of the sugar beet rhizomania disease. The virus is currently listed in Annex III as a protected zone (PZ) quarantine pest of the Commission Implementing Regulation (EU) 2019/2072. The identity of the BNYVV is well established. BNYVV is a soil-borne virus transmitted by the obligate root plasmodiophorid endoparasite Polymyxa betae. BNYVV is widely distributed in the EU, but is not reported in the following EU PZs: Ireland, France (Brittany), Portugal (Azores), Finland and Northern Ireland. The virus may enter, become established and spread in the PZs via P. betae resting spores with soil and growing media as such or attached to machinery and with roots and tubercles of species other than B. vulgaris and with plants for planting. Introduction of BNYVV would have a negative impact on sugar beet and other beet crops in PZs, because of yield and sugar content reduction. Phytosanitary measures are available to reduce the likelihood of entry and spread in the PZs. Once the virus and its plasmodiophorid vector have entered a PZ, their eradication would be difficult due to the persistence of viruliferous resting spores in the soil. The main knowledge gaps or uncertainties identified concerning the presence of BNYVV in the PZs and the incidence and distribution of BNYVV in Switzerland, a country to which a range of specific requirements do not apply. BNYVV meets all the criteria that are within the remit of EFSA to qualify as a potential protected zone union quarantine pest. Plants for planting are not considered as a main means of spread, and therefore BNYVV does not satisfy all the criteria evaluated by EFSA to qualify as potential Union regulated non-quarantine pest.


Dynamical scaling of imbibition in columnar geometries.
Recent experiments on imbibition in columnar geometries show interfacial fluctuations whose dynamic scaling is not compatible with the usual nonlocal model governed by surface tension that results from a macroscopic description. To explore this discrepancy, we exhaustively analyze numerical integrations of a phase-field model with dichotomic columnar disorder. We find that two distinct behaviors are possible depending on the capillary contrast between the two values of disorder. In a high-contrast case, where interface evolution is mainly dominated by the disorder, an inherent anomalous scaling is always observed. Moreover, in agreement with experimental work, the interface motion has to be described through a local model. On the other hand, in a lower-contrast case, the interface is dominated by interfacial tension and can be well modeled by a nonlocal model. We have studied both spontaneous and forced-flow imbibition situations, giving a complete set of scaling exponents in each case, as well as a comparison to the experimental results.


Is EuroSCORE II better than EuroSCORE in predicting mortality after transcatheter aortic valve implantation?
The purpose of this study was to examine the performance of the European system for cardiac operative risk evaluation II (ESII) in patients undergoing transcatheter aortic valve implantation (TAVI). The ESII was developed recently to improve the predictive value of the original logistic EuroSCORE (LES). Between October 2006 and November 2011, 453 consecutive patients with severe aortic stenosis undergoing TAVI with either the Edwards valve or the Corevalve were included in the current analysis. The performance of the ESII, LES, and society of thoracic surgeons predicted risk of mortality score (STS) was evaluated. Mean age was 83.1 ± 6.4 years. The Edwards valve was used in 382 patients (84.3%) of the cohort, transfemoral approach (TF) in 55.0%, transapical approach (TA) in 25.2%, transaortic approach (TAo) 17.8%, transsubclabian approach (SC) 2.0%. The observed 30-day mortality was 12.6% (11.2, 18.4, 7.4, and 22.2% for TF, TA, TAo, SC, respectively). The mean LES, STS, and ESII were 22.4 ± 12.1, 8.1 ± 6.0, and 8.1 ± 5.2, respectively. The Hosmer-Lemeshow test showed ESII was inadequately calibrated for 30-day mortality compared with other risk scores (ESII P = 0.09, LES P = 0.84, STS P = 0.34). By using the area under the receiver operating characteristic curve (AUC), ESII better predicted 30-day mortality, albeit poorly, compared to LES and STS (AUC = 0.68, 0.65, and 0.60, respectively). In the TF cohort, ESII was better in predicting 30-day mortality compared to LES and STS (AUC = 0.74, 0.61, 0.60, respectively). Although the ESII demonstrated better predictive performance especially in the TF cohort, ESII is still inadequate in predicting mortality after TAVI as are LES and STS.


The mitotic apparatus: isolation by controlled pH.
The isolation of the mitotic apparatus (MA) from the echinoderm egg was studied in detail, with particular attention given to the factors governing its stability. Successful isolation depends mainly on the pH of the isolation solution, slightly acid values being required. The use of a 1 M solution of hexanediol, buffered at pH 6.0 to 6.4, gives high yields of stable MA, while MA of poorer quality can be isolated in water buffered at pH 5.5 to 5.8. Isolation is possible only over a very narrow range of pH, as the cells become more difficult to break at lower values and the MA becomes unstable at higher values. Within this range the fibrous structure of the MA varies with the pH. The isolated MA disintegrates slowly when transferred to water at pH 7 and dissolves rapidly in solutions of high ionic strength.


Characteristics and outcome among patients with a suspected in-hospital cardiac arrest.
To describe the characteristics and outcome among patients with a suspected in-hospital cardiac arrest. All the patients who suffered from a suspected in-hospital cardiac arrest during a 14-months period, where the cardiopulmonary resuscitation (CPR) team was called, were recorded and described prospectively in terms of characteristics and outcome. There were 278 calls for the CPR team. Of these, 216 suffered a true cardiac arrest, 16 a respiratory arrest and 46 neither. The percentage of patients who were discharged alive from hospital was 42% for cardiac arrest patients, 62% for respiratory arrest and 87% for the remaining patients. Among patients with a cardiac arrest, those found in ventricular fibrillation/ventricular tachycardia had a survival rate of 64%, those found in asystole 24% and those found in pulseless electrical activity 10%. Among patients who were being monitored at the time of arrest, the survival rate was 52%, as compared with 27% for non-monitored patients (P= 0.001). Among survivors of cardiac arrest, a cerebral performance category (CPC) of 1 (no major deficit) was observed in 81% at discharge and in 82% on admission to hospital prior to the arrest. We conclude that, during a 14-month period at Sahlgrenska University Hospital in Göteborg, almost half the patients with a cardiac arrest in which the CPR team was called were discharged from hospital. Among survivors, 81% had a CPC score of 1 at hospital discharge. Survival seems to be closely related to the relative effectiveness of the resuscitation organisation in different parts of the hospital.


An Extension of the Arden Syntax to Facilitate Clinical Document Generation.
While clinical information systems usually store patient records in database tables, human interpretations as well as information transfer between institutions often require that clinical data can be represented as documents. To automate document generation from patient data in conjunction with the rich computational facilities of clinical decision support, we propose a template-based extension of the Arden Syntax, and discuss the benefits and limitations observed during a pilot application for patient recruitment. While the original Arden Syntax supports string concatenation as well as the substitution of unnamed placeholders, we integrated an additional method based on embedding expressions into strings. A dedicated parser identifies the expressions and automatically substitutes them at runtime, which can for example be harnessed to display the most recent value from a time series. The resulting mechanism supports the generation of extensive clinical documents without the need to apply specific operators. To evaluate the proposed extension, we implemented an Arden module that identifies an intensive care patient cohort that conforms to the eligibility criteria of a clinical trial and outputs a concise patient overview in different document formats. While string interpolation in the original Arden standard has been tailored to clinical event monitoring, we interpret that our accessible approach usefully extends Arden's data-to-text capabilities. Future research might target the development of an interactive template editor that would hide the complexity of formatting directives and conditional expressions behind a graphical user interface, and explore how computer-linguistic formalisms might facilitate advanced features such as automatic inflections of verbs and nouns.


State-level structural sexual stigma and HIV prevention in a national online sample of HIV-uninfected MSM in the United States.
Stigmatizing social environments (of which 'structural stigma' is one component) negatively affect health-related outcomes. However, few studies have examined structural stigma related to sexual minority status as a risk factor for HIV outcomes among MSM. In August 2013, members of a large MSM social and sexual networking site in the United States completed a survey about HIV-prevention practices. A previously validated composite index provided values for state-level structural stigma, including density of same-sex couples, proportion of public high schools with Gay-Straight Alliances, state laws protecting sexual minorities, and public opinion toward homosexuality. Multivariable logistic generalized estimating equations assessed the relationship between structural stigma and condomless anal intercourse, use and awareness of antiretroviral-based HIV-prevention strategies (i.e. pre and postexposure prophylaxis, or PEP and PrEP), and comfort discussing male-male sex with primary care providers. Among the 4098 HIV-uninfected MSM, lower state-level structural stigma was associated with decreased odds of condomless anal intercourse [adjusted odds ratio (aOR) 0.97 per one unit increase in structural stigma score, 95% confidence interval (CI) 0.94-0.99], increased odds of awareness of PEP (aOR 1.06, 95% CI 1.02-1.09), and PrEP (aOR 1.06, 95% CI 1.02-1.10), having taken PEP (aOR 1.15, 95% CI 1.05-1.26) and PrEP (aOR 1.21, 95% CI 1.01-1.44), and comfort discussing male-male sex with providers (aOR 1.08, 95% CI 1.05-1.11), after adjusting for social and state-level confounders. MSM living in more stigmatizing environments had decreased use of antiretroviral-based HIV-prevention strategies compared to those in less stigmatizing environments. Legal reforms protecting sexual minorities should be evaluated as structural interventions that could reduce HIV risk among MSM.


Structural analysis of the interaction between the SNARE Tlg1 and Vps51.
Membrane fusion in cells involves the interaction of SNARE proteins on apposing membranes. Formation of SNARE complexes is preceded by tethering events, and a number of protein complexes that are thought to mediate this have been identified. The VFT or GARP complex is required for endosome-Golgi traffic in yeast. It consists of four subunits, one of which, Vps51, has been shown to bind specifically to the SNARE Tlg1, which participates in the same fusion event. We have determined the structure of the N-terminal domain of Tlg1 bound to a peptide from the N terminus of Vps51. Binding depends mainly on residues 18-30 of Vps51. These form a short helix which lies in a conserved groove in the three-helix bundle formed by Tlg1. Surprisingly, although both Vps51 and Tlg1 are required for transport to the late Golgi from endosomes, removal of the Tlg1-binding sequences from Vps51 does not block such traffic in vivo. Thus, this particular interaction cannot be crucial to the process of vesicle docking or fusion.


Effectiveness of a Domestic Abuse Program for Australian Indigenous Offenders.
The subject of this study is to assess the effectiveness of a culture-neutral domestic abuse program (DAP) developed for offenders with domestic and family violence histories, when used for Australian Indigenous offenders, serving community-based supervised orders. The study employed a retrospective quasi-experimental research design and intention-to-treat program definition for 953 DAP-treated men and propensity score-matched controls, from diverse cultural, religious, and racial backgrounds, 19% being Indigenous Australians. Outcomes measured were program completion, time to first reconviction, and reconviction rates. Indigenous men completed the program similar to non-Indigenous men (58% vs. 63%; NS), although significantly more dropped out (22% vs. 18%); 63% of Indigenous DAP-treated men, remained reconviction free versus 49% of Indigenous controls. Significant therapeutic benefits required program completion, 73% Indigenous and 74% of non-Indigenous men remained reconviction free. Indigenous DAP participants, relative to controls, took significantly longer to first reconviction. Survival was associated with prior criminal histories, but not with rural or remote domiciles. Reconviction rates were predicted by Level of Service Inventory-Revised actuarial risk scores and by DAP completion, but not by Indigenous status. Program effect size was d = .477, mean reconvictions for Indigenous DAP enrollees being 50% lower than controls. Generic domestic violence interventions utilising evidence-based theoretical principles can be effective for Indigenous offenders, despite an absence of specific Indigenous cultural, or healing content, and delivery by Indigenous facilitators or Elders.


Ethnopharmacological field studies: a critical assessment of their conceptual basis and methods.
Ethnopharmacology as a well-defined field has a relatively short history, but for centuries researchers have been interested in the observation, description, and experimental investigation of indigenous drugs and their biological activities. Today, such articles are published in a variety of journals among which the Journal of Ethnopharmacology has a prominent position as well as in book monographs. As any other area of scientific endeavour, this field requires a critical and engaged discussion about the conceptual basis, the relevant methods and the overall standards necessary for excellence. Here we review recent ethnopharmacological field studies in order to highlight achievements and future needs for improving the quality of such studies. The basis for this review is 40 field studies published in the years 2007 and 2008 in the Journal of Ethnopharmacology. Researchers need to have a clear vision for what and how they want to achieve a conceptually and methodologically sound approach and as in all disciplines adherence to internationally recognized methodological standards is essential. Here we review not only the basic conceptual requirements but also the minimal methodological (i.e. botanical, anthropological/historical,ethnomedical) standards and ways how to quantify ethnopharmacological information. Future uses of such information both in the context of experimental research and in applied projects highlight the multiple roles of such data generated in ethnopharmacological field studies. This review cannot be a book of recipes on how to conduct such research but highlights minimal conceptual and methodological requirements for use in future projects.


Comparative safety and effectiveness of alendronate versus raloxifene in women with osteoporosis.
Alendronate and raloxifene are among the most popular anti-osteoporosis medications. However, there is a lack of head-to-head comparative effectiveness studies comparing the two treatments. We conducted a retrospective large-scale multicenter study encompassing over 300 million patients across nine databases encoded in the Observational Medical Outcomes Partnership (OMOP) Common Data Model (CDM). The primary outcome was the incidence of osteoporotic hip fracture, while secondary outcomes were vertebral fracture, atypical femoral fracture (AFF), osteonecrosis of the jaw (ONJ), and esophageal cancer. We used propensity score trimming and stratification based on an expansive propensity score model with all pre-treatment patient characteritistcs. We accounted for unmeasured confounding using negative control outcomes to estimate and adjust for residual systematic bias in each data source. We identified 283,586 alendronate patients and 40,463 raloxifene patients. There were 7.48 hip fracture, 8.18 vertebral fracture, 1.14 AFF, 0.21 esophageal cancer and 0.09 ONJ events per 1,000 person-years in the alendronate cohort and 6.62, 7.36, 0.69, 0.22 and 0.06 events per 1,000 person-years, respectively, in the raloxifene cohort. Alendronate and raloxifene have a similar hip fracture risk (hazard ratio [HR] 1.03, 95% confidence interval [CI] 0.94-1.13), but alendronate users are more likely to have vertebral fractures (HR 1.07, 95% CI 1.01-1.14). Alendronate has higher risk for AFF (HR 1.51, 95% CI 1.23-1.84) but similar risk for esophageal cancer (HR 0.95, 95% CI 0.53-1.70), and ONJ (HR 1.62, 95% CI 0.78-3.34). We demonstrated substantial control of measured confounding by propensity score adjustment, and minimal residual systematic bias through negative control experiments, lending credibility to our effect estimates. Raloxifene is as effective as alendronate and may remain an option in the prevention of osteoporotic fracture.


Changes in volatile production during the course of fungal mycelial interactions between Hypholoma fasciculare and Resinicium bicolor.
The mycelia of two wood decay basidiomycete fungi were grown opposing each other across a 1-microm pore membrane supported on the surface of malt broth, contained within a sealable reaction vessel. Production of volatiles during the time course of interaction was followed by collecting head space samples by solid phase microextraction (100 microm polydimethylsiloxane fiber) on five occasions over 25 d following coinoculation of the fungi: 1, 3 (i.e., immediately prior to mycelial contact), 9 (1-2 d after initiation of pigment production by Resinicium bicolor), 17, and 25 d. Ten volatiles were produced during interactions that were not detected in single species controls. In general, most (18) fungal volatiles were sesquiterpenes eluted between 12.5 and 21 min, with a further two eluted at 29.1 and 33.9 min; a benzoic acid methyl ester, a benzyl alcohol, and a quinolinium type compound with a distinctive fragmentation pattern at m/z 203, 204, 206, and 207 were also identified; three volatiles with m/z maxima of 163, 159, and 206-208, respectively, remained unidentified. The results are discussed in relation to possible ecological roles of volatiles.


Microscope calibration using laser written fluorescence.
There is currently no widely adopted standard for the optical characterization of fluorescence microscopes. We used laser written fluorescence to generate two- and three-dimensional patterns to deliver a quick and quantitative measure of imaging performance. We report on the use of two laser written patterns to measure the lateral resolution, illumination uniformity, lens distortion and color plane alignment using confocal and structured illumination fluorescence microscopes.


Myo10 is a key regulator of TNT formation in neuronal cells.
Cell-to-cell communication is essential in multicellular organisms. Tunneling nanotubes (TNTs) have emerged as a new type of intercellular spreading mechanism allowing the transport of various signals, organelles and pathogens. Here, we study the role of the unconventional molecular motor myosin-X (Myo10) in the formation of functional TNTs within neuronal CAD cells. Myo10 protein expression increases the number of TNTs and the transfer of vesicles between co-cultured cells. We also show that TNT formation requires both the motor and tail domains of the protein, and identify the F2 lobe of the FERM domain within the Myo10 tail as necessary for TNT formation. Taken together, these results indicate that, in neuronal cells, TNTs can arise from a subset of Myo10-driven dorsal filopodia, independent of its binding to integrins and N-cadherins. In addition our data highlight the existence of different mechanisms for the establishment and regulation of TNTs in neuronal cells and other cell types.


Derivation of ganglioside-specific T cell lines of suppressor or helper phenotype from cerebrospinal fluid of multiple sclerosis patients.
In order to investigate the specificity of activated T cells in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS), we have cultured cells in the presence of mitogen-free IL-2 but without any antigen. Two T cell lines have been derived and showed specific reactivity to certain purified gangliosides (GM1, GD1a, GD1b and GQ1b). However, responses to other brain and viral antigens were not seen, and neither were T cell lines from peripheral blood lymphocytes (PBL) of normal, MS or other neurological disease patients stimulated by these gangliosides. Release of IL-2 could be detected after incubation of these CSF lines with specific gangliosides. One line exhibited predominantly helper/inducer (T4+) phenotype whilst the other was suppressor/cytotoxic (T8+), and further analysis indicated it could be of the suppressor phenotype. These data may have implications for T cell-induced demyelination in MS.


The currency, completeness and quality of systematic reviews of acute management of moderate to severe traumatic brain injury: A comprehensive evidence map.
To appraise the currency, completeness and quality of evidence from systematic reviews (SRs) of acute management of moderate to severe traumatic brain injury (TBI). We conducted comprehensive searches to March 2016 for published, English-language SRs and RCTs of acute management of moderate to severe TBI. Systematic reviews and RCTs were grouped under 12 broad intervention categories. For each review, we mapped the included and non-included RCTs, noting the reasons why RCTs were omitted. An SR was judged as 'current' when it included the most recently published RCT we found on their topic, and 'complete' when it included every RCT we found that met its inclusion criteria, taking account of when the review was conducted. Quality was assessed using the AMSTAR checklist (trichotomised into low, moderate and high quality). We included 85 SRs and 213 RCTs examining the effectiveness of treatments for acute management of moderate to severe TBI. The most frequently reviewed interventions were hypothermia (n = 17, 14.2%), hypertonic saline and/or mannitol (n = 9, 7.5%) and surgery (n = 8, 6.7%). Of the 80 single-intervention SRs, approximately half (n = 44, 55%) were judged as current and two-thirds (n = 52, 65.0%) as complete. When considering only the most recently published review on each intervention (n = 25), currency increased to 72.0% (n = 18). Less than half of the 85 SRs were judged as high quality (n = 38, 44.7%), and nearly 20% were low quality (n = 16, 18.8%). Only 16 (20.0%) of the single-intervention reviews (and none of the five multi-intervention reviews) were judged as current, complete and high-quality. These included reviews of red blood cell transfusion, hypothermia, management guided by intracranial pressure, pharmacological agents (various) and prehospital intubation. Over three-quarters (n = 167, 78.4%) of the 213 RCTs were included in one or more SR. Of the remainder, 17 (8.0%) RCTs post-dated or were out of scope of existing SRs, and 29 (13.6%) were on interventions that have not been assessed in SRs. A substantial number of SRs in acute management of moderate to severe TBI lack currency, completeness and quality. We have identified both potential evidence gaps and also substantial research waste. Novel review methods, such as Living Systematic Reviews, may ameliorate these shortcomings and enhance utility and reliability of the evidence underpinning clinical care.


Difference in dental lateral arch length between 9-year-olds born in the 1960s and the 1980s.
This study was carried out in two cohorts of 9-year-olds, born in the 1960s and the 1980s in Sweden and Norway, to test the hypothesis that there has been a change in the lateral arch space conditions of the dentition during the last decades. In Norway, 61 children born in 1963 and 58 children born in 1983 were examined. In Sweden, 66 children born in 1961 and 67 children born in 1984 were examined. Children with a prolonged sucking habit, major dentofacial anomalies, history of orthodontic treatment, and ancestors of foreign origin were excluded to avoid influence of these factors on the development of the dentition. Linear measurements were performed between the mesial surface of the first permanent molars and the distal surface of the permanent lateral incisors in the maxillary and mandibular dental arches with use of a sliding caliper. The results showed that the children born in the 1980s had significantly larger lateral arch spaces in the maxillary dental arch (P <.05) and in the mandibular dental arch (P <.01) than did the children born in the 1960s. The amount of tissue loss in the second deciduous molars (the teeth having caries lesions or having been extracted) was greater in the 1960s groups than in the 1980s groups (P <.001). Thus, the hypothesis, that there has been a change in the lateral arch space conditions during the last decades, was confirmed. The most obvious reason is the decline in caries prevalence that has occurred in the same period; thus, there has been less loss of proximal tooth material of posterior teeth.


Ultrasmall spot size scanning laser ophthalmoscopy.
An ultrasmall spot size scanning laser ophthalmoscope has been developed that employs an annular aberration-corrected incident beam to increase the effective numerical aperture of the eye thereby reducing the width of the probing light spot. Parafovea and foveal cone photoreceptor visibility determined from small area retinal image scans are discussed from the perspective of mode matching between the focused incident beam and the waveguide modes of individual cones. The cone visibility near the fovea centralis can be increased with the annular illumination scheme whereas the visibility of larger parafovea cones drops significantly as a consequence of poorer mode match. With further improvements of the implemented wavefront correction technology it holds promise for individual cone-photoreceptor imaging at the fovea centralis and for optical targeting of the retina with increased resolution.


The ubiquitin E3 ligase LOSS OF GDU2 is required for GLUTAMINE DUMPER1-induced amino acid secretion in Arabidopsis.
Amino acids serve as transport forms for organic nitrogen in the plant, and multiple transport steps are involved in cellular import and export. While the nature of the export mechanism is unknown, overexpression of GLUTAMINE DUMPER1 (GDU1) in Arabidopsis (Arabidopsis thaliana) led to increased amino acid export. To gain insight into GDU1's role, we searched for ethyl-methanesulfonate suppressor mutants and performed yeast-two-hybrid screens. Both methods uncovered the same gene, LOSS OF GDU2 (LOG2), which encodes a RING-type E3 ubiquitin ligase. The interaction between LOG2 and GDU1 was confirmed by glutathione S-transferase pull-down, in vitro ubiquitination, and in planta coimmunoprecipitation experiments. Confocal microscopy and subcellular fractionation indicated that LOG2 and GDU1 both localized to membranes and were enriched at the plasma membrane. LOG2 expression overlapped with GDU1 in the xylem and phloem tissues of Arabidopsis. The GDU1 protein encoded by the previously characterized intragenic suppressor mutant log1-1, with an arginine in place of a conserved glycine, failed to interact in the multiple assays, suggesting that the Gdu1D phenotype requires the interaction of GDU1 with LOG2. This hypothesis was supported by suppression of the Gdu1D phenotype after reduction of LOG2 expression using either artificial microRNAs or a LOG2 T-DNA insertion. Altogether, in accordance with the emerging bulk of data showing membrane protein regulation via ubiquitination, these data suggest that the interaction of GDU1 and the ubiquitin ligase LOG2 plays a significant role in the regulation of amino acid export from plant cells.


Housing and location choices of retiring households: evidence from France.
In this paper, a study is made of the mobility and housing choices of the elderly when retiring, using household data collected in France. From a theoretical viewpoint, individuals are likely to decrease their housing quantity because of an income loss when retiring, but they may also increase it to benefit from more housing comfort for leisure. Using the 1992 Trois Générations survey, it is first shown that housing mobility at retirement is substantial in France, with a variety of self-reported motives. Then, using the 1994—2001 French Europanel survey, evidence is found of both upsizing and downsizing for mobile recent retirees. In many cases, housing adjustments lead to a correction of the initial disequilibrium between the number of rooms and the number of occupants. However, a significant proportion of mobile recent retirees improve the quality of their dwelling.


Rationale and design of the MEMA kwa Vijana adolescent sexual and reproductive health intervention in Mwanza Region, Tanzania.
Large-scale innovative, integrated, multifaceted adolescent sexual and reproductive health (ASRH) interventions are urgently needed in sub-Saharan Africa. Implementation through schools and health facilities may maximize intervention coverage and sustainability, however the impact of the use of these structures on intervention content and delivery is not well documented. This paper describes the rationale and design of a large-scale multifaceted ASRH intervention, which was developed and evaluated over three years in rural communities in Mwanza Region, North West Tanzania. The intervention comprised community mobilization, participatory reproductive health education in primary schools, youth-friendly reproductive health services and community-based condom provision for youth. We examine the effect of socioeconomic, cultural and infrastructural factors on intervention content and implementation. This paper demonstrates the means by which such interventions can be feasibly and sustainably implemented to a high standard through existing government health and school structures. However, the use of these structures involves compromise on some key aspects of intervention design and requires the development of complementary strategies to access out-of-school youth and the wider community.


Stability and prognostic value of Slug, Sox9 and Sox10 expression in breast cancers treated with neoadjuvant chemotherapy.
Expression of transcription-factors as Slug and Sox9 was recently described to determine mammary stem-cell state. Sox10 was previously shown to be present also in breast cancer. Protein overexpression of Slug, Sox9 and Sox10 were associated with poor overall survival and with triple-negative phenotype in breast cancer. In this study we tested the stability of Slug, Sox9 and Sox10 expression during chemotherapy and addressed their prognostic role of in neoadjuvant treated primary breast-cancer and their correlation to pathological-response and overall survival. We analyzed immunohistochemical expression of Slug, Sox9 and Sox10 in tissue microarrays of 96 breast cancers prior to and after neoadjuvant chemotherapy. Expression was evaluated in invasive tumor cells and in tumor stroma and scored as 0, 1+, 2+ 3+. Expression-profile prior to and after chemotherapy was correlated to overall survival (Kaplan Meier) and with established clinico-pathological parameter. Sox9, Sox10 and Slug were expressed in 82-96% of the tumor cells prior to chemotherapy. Slug was expressed in 97% of the cases in tumor stroma before therapy. Change in expression-profile after chemotherapy occurred only in Slug expression in tumor-cells (decreased from 82 to 51%, p = 0.0001, Fisher's exact test). The other markers showed no significant change after chemotherapy. Stromal Sox9 expression (0 to 2+) correlated to better overall survival after chemotherapy (p = 0.004) and reached almost statistical significance prior to chemotherapy (p = 0.065). There was no correlation between Sox9 and hormone-receptor expression. In multivariate-analysis, the stromal Sox9 expression after chemotherapy proved to be an independent and better prognostic marker than hormone-receptor status. Other clinico-pathological parameter (as HER2-status or pathological-stage) showed no correlation to the analyzed markers. Strong stromal Sox9 expression in breast cancer after chemotherapy was found to bear negative prognostic information and was associated with shortened overall survival. Slug expression was significantly changed (reduced) in samples after neoadjuvant chemotherapy.


Virion RNA species of the arenaviruses Pichinde, Tacaribe, and Tamiami.
The principal RNA species isolated from labeled preparations of the arenavirus Pichinde usually include a large viral RNA species L (apparent molecular weight = 3.2 X 10(6)), and a smaller viral RNA species S (apparent molecular weight = 1.6 X 10(6)). In addition, either little or considerable quantities of 28S rRNA as well as 18S rRNA can also be obtained in virus extracts, depending on the virus stock and growth conditions used to generate virus preparations. Similar RNA species have been identified in RNA extracted from Tacaribe and Tamiami arenavirus preparations. Oligonucleotide fingerprint analyses have confirmed the host ribosomal origin of the 28S and 18S species. Such analyses have also indicated that the Pichinde viral L and S RNA species each contain unique nucleotide sequences. Viral RNA preparations isolated by conventional phenol-sodium dodecyl sulfate extraction often have much of their L and S RNA species in the form of aggregates as visualized by either electron microscopy or oligonucleotide fingerprinting of material recovered from the top of gels (run by using undenatured RNA preparations). Circular and linear RNA forms have also been seen in electron micrographs of undenatured RNA preparations, although denatured viral RNA preparations have yielded mostly linear RNA species with few RNA aggregates or circular forms.


Reappraisal of the diuretic effect of empagliflozin in the EMPA-REG OUTCOME trial: Comparison with classic diuretics.
Empagliflozin, a sodium-glucose cotransporter type 2 (SGLT2) inhibitor, has been associated with a remarkable reduction in cardiovascular and all-cause mortality in patients with type 2 diabetes and antecedents of cardiovascular disease. This effect was attributed to a diuretic (haemodynamic) rather than metabolic (antiatherogenic) effect. The aim of this review is to offer arguments that either support or challenge this 'diuretic hypothesis'. The literature was scrutinized to: (1) examine the diuretic effects of SGLT2 inhibitors vs. hydrochlorothiazide as the reference diuretic; (2) analyze the effects of classic diuretics on cardiovascular outcomes and mortality in diabetic patients; and (3) reconsider some of the specific analyses of the EMPA-REG OUTCOME trial possibly related to a diuretic effect. The diuretic effect of empagliflozin has so far been poorly investigated, although SGLT2 inhibitors have actions distinct from those of classic diuretics. The effects of thiazide-like diuretics on cardiovascular and overall mortality have been limited in diabetic patients with hypertension, whereas the effects of mineralocorticoid receptor antagonists in subgroups of diabetic patients with heart failure were more impressive, but still largely inferior to those reported in EMPA-REG, where relative reductions in mortality with empagliflozin were observed in diabetic patients with or without heart failure, arterial hypertension, renal impairment or diuretic background therapy. Although the diuretic hypothesis was put forward to explain the remarkable reduction in mortality with empagliflozin in EMPA-REG, the available results do not support a major contribution of this mechanism, unless the specific diuretic effect of SGLT2 inhibitors turns out to be markedly different from those of classic diuretics.


Decision aids for people facing health treatment or screening decisions.
Decision aids prepare people to participate in preference-sensitive decisions. 1. Create a comprehensive inventory of patient decision aids focused on healthcare options. 2. Review randomized controlled trials (RCT) of decision aids, for people facing healthcare decisions. Studies were identified through databases and contact with researchers active in the field. Two independent reviewers screened abstracts for interventions designed to aid patients' decision making by providing information about treatment or screening options and their associated outcomes. Information about the decision aids was compiled in an inventory; those that had been evaluated in a RCT were reviewed in detail. Two reviewers independently extracted data using standardized forms. Results of RCTs were pooled using weighted mean differences (WMD) and relative risks (RR) using a random effects model. Over 200 decision aids were identified. Of the 131 available decision aids, most are intended for use before counselling. Using the CREDIBLE criteria to evaluate the quality of the decision aids: a) most included potential harms and benefits, credentials of the developers, description of their development process, update policy, and were free of perceived conflict of interest; b) many included reference to relevant literature; c) few included a description of the level of uncertainty regarding the evidence; and d) few were evaluated. Thirty of these decision aids were evaluated in 34 RCTs and another trial evaluated a suite of eight decision aids. An additional 30 trials are yet to be published. Among the trials comparing decision aids to usual care, decision aids performed better in terms of: a) greater knowledge (WMD 19 out of 100, 95% CI: 13 to 24; b) more realistic expectations (RR 1.4, 95%CI: 1.1 to 1.9); c) lower decisional conflict related to feeling informed (WMD -9.1 of 100, 95%CI: -12 to -6); d) increased proportion of people active in decision making (RR 1.4, 95% CI: 1.0 to 2.3); and e) reduced proportion of people who remained undecided post intervention (RR 0.43, 95% CI: 0.3 to 0.7). When simpler were compared to more detailed decision aids, the relative improvement was significant in: a) knowledge (WMD 4 out of 100, 95% CI: 3 to 6); b) more realistic expectations (RR 1.5, 95% CI: 1.3 to 1.7); and c) greater agreement between values and choice. Decision aids appeared to do no better than comparisons in affecting satisfaction with decision making, anxiety, and health outcomes. Decision aids had a variable effect on which healthcare options were selected. The availability of decision aids is expanding with many on the Internet; however few have been evaluated. Trials indicate that decision aids improve knowledge and realistic expectations; enhance active participation in decision making; lower decisional conflict; decrease the proportion of people remaining undecided, and improve agreement between values and choice. The effects on persistence with chosen therapies and cost-effectiveness require further evaluation. Finally, optimal strategies for dissemination need to be explored.


Dose adjustment of carboplatin in patients on hemodialysis.
Carboplatin is one of the most prescribed cytotoxic drug, which is extensively used in the treatment regimens of several malignancies. The therapeutic efficiency of carboplatin has been found to correlate the area under curve (AUC). The Calvert formula has been extensively used to determine the dose of carboplatin for a fixed AUC and glomerular filtration rate (GFR). This formula has also been used in patients with end-stage renal disease on hemodialysis by assuming that the GFR is zero. This is applicable to patients who receive hemodialysis within 12-18 h after carboplatin infusion. After the first 24 h, a majority of the carboplatin is bound to proteins is not easily dialyzable and hence continues to remain in the blood stream despite repeated sessions of hemodialysis. We derive a correction factor to calculate the resultant AUC in such patients. The analysis done by using this correction factor shows that the AUC can increase by eightfold in patients who received the adjusted dose but whose hemodialysis was delayed beyond 24 h after infusion. The correction factor proposed here can also be used to calculate the dose adjustment required a priori in patients who may receive delayed hemodialysis. It is also useful to predict the AUC and estimate the resultant toxicity in such patients.


An adenosine-mediated signaling pathway suppresses prenylation of the GTPase Rap1B and promotes cell scattering.
During metastasis, cancer cells acquire the ability to dissociate from each other and migrate, which is recapitulated in vitro as cell scattering. The small guanosine triphosphatase (GTPase) Rap1 opposes cell scattering by promoting cell-cell adhesion, a function that requires its prenylation, or posttranslational modification with a carboxyl-terminal isoprenoid moiety, to enable its localization at cell membranes. Thus, signaling cascades that regulate the prenylation of Rap1 offer a mechanism to control the membrane localization of Rap1. We identified a signaling cascade initiated by adenosine A2B receptors that suppressed the prenylation of Rap1B through phosphorylation of Rap1B, which decreased its interaction with the chaperone protein SmgGDS (small GTPase guanosine diphosphate dissociation stimulator). These events promoted the cytosolic and nuclear accumulation of nonprenylated Rap1B and diminished cell-cell adhesion, resulting in cell scattering. We found that nonprenylated Rap1 was more abundant in mammary tumors than in normal mammary tissue in rats and that activation of adenosine receptors delayed Rap1B prenylation in breast, lung, and pancreatic cancer cell lines. Our findings support a model in which high concentrations of extracellular adenosine, such as those that arise in the tumor microenvironment, can chronically activate A2B receptors to suppress Rap1B prenylation and signaling at the cell membrane, resulting in reduced cell-cell contact and promoting cell scattering. Inhibiting A2B receptors may be an effective method to prevent metastasis.


Differential inhibition of thrombin generation by vitamin K antagonists alone and associated with low-molecular-weight heparin.
Vitamin K antagonists (VKA) treatment starts with co-administration of low-molecular-weight heparin (LMWH). The anticoagulation induced by the two drugs is still not well determined. In the present study we used thrombin generation assay to evaluate the hypo-coagulation induced by treatment with VKA and by the combination of VKA with LMWH. Tissue factor triggered thrombin generation in platelet-poor plasma was assessed in samples from 15 healthy volunteers, 97 samples from patients treated with VKA and 41 samples from patients receiving enoxaparin and VKA. Patients were classified according to international normalised ratio (INR) level (<2, 2-3 and >3). In plasma samples from patients treated with VKA having INR 2-3 the inhibition of thrombin generation reached 50% compared to controls. In samples with INR>3 this inhibition was 80%. In samples from patients receiving both LMWH and VKA, thrombin generation was significantly decreased compared to the controls and VKA group. In samples with an INR 2-3 obtained from patients treated with LMWH and VKA, the inhibition of thrombin generation was similar to that observed in samples with an INR>3 obtained from VKA treated patients. Thrombin generation assay is sensitive to detect the global the anticoagulant effect produced by the association of LMWH and VKA. For equal INR dual anticoagulant treatment induces significantly more profound inhibition of thrombin generation compared to treatment with VKA alone. The clinical relevance of this observation merits to be studied in prospective studies in patients with defined indications of anticoagulant therapy.


Effects of the mutation R145G in human cardiac troponin I on the kinetics of the contraction-relaxation cycle in isolated cardiac myofibrils.
Familial hypertrophic cardiomyopathy (FHC) has been linked to mutations in sarcomeric proteins such as human cardiac troponin I (hcTnI). To elucidate the functional consequences of the mutation hcTnI(R145G) on crossbridge kinetics, force kinetics were analysed in murine cardiac myofibrils carrying either the mutant or the wild-type protein. The mutation was introduced into the myofibrils in two different ways: in the first approach, the endogenous Tn was replaced by incubation of the myofibrils with an excess of reconstituted recombinant hcTn containing either hcTnI(WT) or hcTnI(R145G). Alternatively, myofibrils were isolated either from non-transgenic or transgenic mice expressing the corresponding mcTnI(R146G) mutation. In myofibrils from both models, the mutation leads to a significant upward shift of the passive force-sarcomere length relation determined at pCa 7.5. Addition of 5 mm BDM (2,3-butandione-2-monoxime), an inhibitor of actomyosin ATPase partially reverses this shift, suggesting that the mutation impairs the normal function of cTnI to fully inhibit formation of force-generating crossbridges in the absence of Ca(2)(+). Maximum force development (F(max)) is significantly decreased by the mutation only in myofibrils exchanged with hcTnI(R145G) in vitro. Ca(2)(+) sensitivity of force development was reduced by the mutation in myofibrils from transgenic mice but not in exchanged myofibrils. In both models the rate constant of force development k(ACT) is reduced at maximal [Ca(2)(+)] but not at low [Ca(2)(+)] where it is rather increased. Force relaxation is significantly prolonged due to a reduction of the relaxation rate constant k(REL). We therefore assume that the impairment in the regulatory function of TnI by the mutation leads to modulations in crossbridge kinetics that significantly alter the dynamics of myofibrillar contraction and relaxation.


Identification of candidate serum proteins for classifying well-differentiated small intestinal neuroendocrine tumors.
Patients with well-differentiated small intestine neuroendocrine tumors (WD-SI-NETs) are most often diagnosed at a metastatic stage of disease, which reduces possibilities for a curative treatment. Thus new approaches for earlier detection and improved monitoring of the disease are required. Suspension bead arrays targeting 124 unique proteins with antibodies from the Human Protein Atlas were used to profile biotinylated serum samples. Discoveries from a cohort of 77 individuals were followed up in a cohort of 132 individuals both including healthy controls as well as patients with untreated primary WD-SI-NETs, lymph node metastases and liver metastases. A set of 20 antibodies suggested promising proteins for further verification based on technically verified statistical significance. Proceeding, we assessed the classification performance in an independent cohort of patient serum, achieving, classification accuracy of up to 85% with different subsets of antibodies in respective pairwise group comparisons. The protein profiles of nine targets, namely IGFBP2, IGF1, SHKBP1, ETS1, IL1α, STX2, MAML3, EGR3 and XIAP were verified as significant contributors to tumor classification. We propose new potential protein biomarker candidates for classifying WD-SI-NETs at different stage of disease. Further evaluation of these proteins in larger sample sets and with alternative approaches is needed in order to further improve our understanding of their functional relation to WD-SI-NETs and their eventual use in diagnostics.


Spatial characteristics of center-surround antagonism in younger and older adults.
Sensitivity to motion direction is affected by stimulus size, contrast (D. Tadin & J. S. Lappin, 2005; D. Tadin, J. S. Lappin, L. A. Gilroy, & R. Blake, 2003), and observer age (L. R. Betts, C. P. Taylor, A. B. Sekuler, & P. J. Bennett, 2005). Here, we investigated the effect of spatial frequency on motion discrimination and how sensitivity changes in older adulthood. We measured stimulus duration thresholds for younger (18-30 years) and older (60-75 years) observers using drifting Gabor gratings that differed in size, spatial frequency, and contrast. A simple model characterized age differences in the threshold-vs.-size functions. The model parameter fits were consistent with an age-related decrease in the strength of spatial suppression. The model also provided good fits to the thresholds when plotted as a function of the total stimulus contrast energy, which suggests that age-related changes in summation and suppression can be modeled as reduced sensitivity to contrast energy. The summation parameter scaled according to stimulus spatial frequency in younger observers only. Suppression strength decreased as a function of spatial frequency in both age groups. Our findings provide additional evidence for age-related changes in summation and suppression mechanisms and suggest that the total contrast energy in the stimulus plays an important role in determining sensitivity to motion direction in both younger and older adults.


Genomic evaluations using data recorded on smallholder dairy farms in low- to middle-income countries.
Breeding has increased genetic gain for dairy cattle in advanced economies but has had limited success in improving dairy cattle in low- to middle-income countries (LMIC). Genetic evaluations are a central component of delivering genetic gain, because they separate the genetic and environmental effects of animals' phenotypes. Genetic evaluations have been successful in advanced economies because of large data sets and strong genetic connectedness, provided by the widespread use of artificial insemination (AI) and accurate recording of pedigree information. In smallholder dairy production systems of many LMICs, the limited use of AI and small herd sizes results in a data structure with insufficient genetic connectedness between herds to facilitate genetic evaluations based on pedigree. Genomic information keeps track of shared haplotypes rather than shared relatives captured by pedigree records. Therefore, genomic information could capture "hidden" genetic relationships, that are not captured by pedigree information, to strengthen genetic connectedness in LMIC smallholder dairy data sets. This study's objective was to use simulation to quantify the power of genomic information to enable genetic evaluation using LMIC smallholder dairy data sets. The results from this study show that (1) genetic evaluations using genomic information were more accurate than those using pedigree information in populations with a high effective population size and weak genetic connectedness; and (2) genetic evaluations modeling herd as a random effect had higher or equal accuracy than those modeling herd as a fixed effect. This demonstrates the potential of genomic information to be an enabling technology in LMIC smallholder dairy production systems by facilitating genetic evaluations with in situ records collected from herds of ≤4 cows. The establishment of routine genomic evaluations could allow the development of LMIC breeding programs comprising an informal set of nucleus animals distributed across many small herds within the target environment. These nucleus animals could be used for genetic evaluation, and the best animals could be disseminated to participating smallholder dairy farms. Together, this could increase the productivity, profitability, and sustainability of LMIC smallholder dairy production systems.